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AB325727

Human IgE ELISA Kit, Chemiluminescent

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Human IgE ELISA Kit, Chemiluminescent is a 55 min single-wash SimpleStep Ignite™ ELISA for the rapid quantification of Human IgE with higher sensitivity and broader measuring range.

- Single 30 min incubation and single -wash step for results in less than an hour
- Chemiluminescent detection allows a broad dynamic range : 10.29 - 22500 pg/ml with a sensitivity of 5.88 pg/mL
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
- Patent pending

View Alternative Names

Immunoglobulin heavy constant epsilon, Ig epsilon chain C region, Ig epsilon chain C region ND, IGHE

6 Images
Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)
  • sELISA

Lab

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)

SimpleStep ELISA® Ignite technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 55 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)
  • sELISA

Lab

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)

Standard curve comparison between the Chemiluminescent Human IgE SimpleStep ELISA and colorimetric Human IgE SimpleStep ELISA (ab317776) in Sample Diluent NS + 1X Enhancer. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)
  • sELISA

Lab

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)

Dynamic range comparison between the Chemiluminescent Human IgE SimpleStep ELISA, colorimetric Human IgE SimpleStep ELISA (ab317776), and leading colorimetric competitor. The Chemiluminescent SimpleStep ELISA kit shows increased sensitivity and increased dynamic range.

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)
  • sELISA

Lab

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)

Interpolated concentration of native IgE was measured in duplicate at different sample concentrations. Undiluted samples are 1 : 40 human serum, plasma (citrate), and plasma (EDTA). The interpolated dilution factor corrected values (to neat samples) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS + 1X Enhancer.

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)
  • sELISA

Lab

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)

Serum of ten individual healthy human female donors was measured in duplicate. Interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). The mean IgE concentration was determined to be 115.5 ng/mL with a range of 25.3 – 303.6 ng/mL.

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)
  • sELISA

Lab

Sandwich ELISA - Human IgE ELISA Kit, Chemiluminescent (AB325727)

Interpolated concentrations of IgE in atopic dermatitis human serum donors. Serum of three individual human donors with atopic dermatitis as measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IgE concentration was determined to be 260.2 ng/mL with a range of 13.9 - 687.2 ng/mL.

Key facts

Detection method

Luminescent

Sample types

Serum, Citrate plasma, EDTA Plasma, Cell culture media, Urine, Saliva

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 5.88 pg/mL

Range

10.29 - 22500 pg/mL

Assay time

55m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Human IgE ELISA Kit, Chemiluminescent (ab325727) is a rapid single-wash sandwich 55 min ELISA to measure Human IgE in Serum, Citrate Plasma, EDTA Plasma, Cell Culture Media, Urine, Saliva with a sensitivity of 5.88 pg/mL.

How the assay works

Human IgE ELISA Kit, Chemiluminescent (ab325727) SimpleStep Ignite™ ELISA employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step during a 30 min only incubation, significantly reducing assay time. By utilizing a chemiluminscent detection method this improves the sensitivity and dynamic range when compared to traditional colormetric ELISAs.

Assay Specificity

Our SimpleStep Ignite™ ELISA kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human IgE (ab325727) protocol summary

1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 30 min at room temperature
2. Wash
3. Add ChemiHRP development solution - incubate for 2 min
4. Read the results on a plate reader capable of luminescence detection

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-IgE antibody [EPR26777-44] - BSA and Azide free (Capture) ab315412
- Anti-IgE antibody [EPR26777-55] - BSA and Azide free (Detector) ab315413

Precision

[ { "reproducibilityType": "Intra", "sample": "Serum", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 2.4" }, { "reproducibilityType": "Inter", "sample": "Serum", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 8.6" } ]

Recovery

[ { "sample": "Serum", "range": "77 - 84 %", "average": "= 81" }, { "sample": "Citrate plasma", "range": "89 - 101 %", "average": "= 97" }, { "sample": "EDTA Plasma", "range": "80 - 88 %", "average": "= 85" }, { "sample": "Cell culture media", "range": "90 - 94 %", "average": "= 93" }, { "sample": "Urine", "range": "98 - 103 %", "average": "= 100" }, { "sample": "Saliva", "range": "93 - 99 %", "average": "= 97" } ]

What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Product protocols

Target data

Constant region of immunoglobulin heavy chains. Immunoglobulins, also known as antibodies, are membrane-bound or secreted glycoproteins produced by B lymphocytes. In the recognition phase of humoral immunity, the membrane-bound immunoglobulins serve as receptors which, upon binding of a specific antigen, trigger the clonal expansion and differentiation of B lymphocytes into immunoglobulins-secreting plasma cells. Secreted immunoglobulins mediate the effector phase of humoral immunity, which results in the elimination of bound antigens (PubMed : 20176268, PubMed : 22158414). The antigen binding site is formed by the variable domain of one heavy chain, together with that of its associated light chain. Thus, each immunoglobulin has two antigen binding sites with remarkable affinity for a particular antigen. The variable domains are assembled by a process called V-(D)-J rearrangement and can then be subjected to somatic hypermutations which, after exposure to antigen and selection, allow affinity maturation for a particular antigen (PubMed : 17576170, PubMed : 20176268).. Isoform 1. Constant region of secreted IgE, also known as the Fc region of IgE antibody. Mediates IgE effector functions on myeloid and lymphoid cells primarily via two Fc receptors, the high-affinity IgE Fc receptor complex/FCER1A : MS4A2 : FCGR1A and the low-affinity FCER2 receptor, which upon antigen/allergen cross-linking initiate signaling pathways that lead to immune cell activation and differentiation (PubMed : 2167225, PubMed : 25629393, PubMed : 33840121, PubMed : 7544003, PubMed : 8114916, PubMed : 8551243). Triggers the immediate hypersensitivity response to allergens as a host defense mechanism against helminth parasites, pathogenic bacteria and venom toxicity. When dysregulated, it can elicit harmful life-threatening allergic and anaphylactic reactions (PubMed : 25629393, PubMed : 33840121, PubMed : 7544003, PubMed : 8114916, PubMed : 8551243). Stimulates the high-affinity IgE Fc receptor complex/FCER1A : MS4A2 : FCGR1A on mast cells, basophils and eosinophils leading to secretion of vasoactive amines, lipid mediators and cytokines that contribute to inflammatory response, tissue remodeling and cytotoxicity against microbes (PubMed : 25629393, PubMed : 8114916, PubMed : 8551243). On macrophages, cross-linking of FCER2 by IgE immune complexes induces intracellular killing of parasites through activation of L-Arginine-nitric oxide pathway (PubMed : 7544003). Activates macrophages to kill tumor cells via antigen-specific antibody-dependent cytotoxicity (ADCC). Triggers differentiation of quiescent M0 macrophages toward M1 state and reprograms M2 macrophages toward a proinflammatory state with antitumor functions (PubMed : 30956175). Stimulates FCER2 on B cells and initiates IgE-dependent antigen uptake and presentation to T cells (PubMed : 2167225).. Isoform 2. Constant region of membrane-bound IgE (long mIgE), part of the B cell receptor complex (BCR). Upon antigen cross-linking triggers quick BCR signaling, ensuring survival of IgE-switched B cells and differentiation into plasma cells, thus regulating both primary and memory IgE responses.. Isoform 3. Constant region of membrane-bound IgE (short mIgE), part of the B cell receptor complex (BCR). Upon antigen cross-linking initiates slower but sustained BCR signaling that negatively regulates mature B cell proliferation.
See full target information IGHE
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